Inhibitory effect of miR-205 targeted YES1 on proliferation of A549 cells
10.13481/j.1671-587x.20140306
- VernacularTitle:miR-205靶定YES1对肺癌细胞A549的增殖抑制作用
- Author:
Yuan CHENG
;
Yongzhan ZHEN
;
Xiaofang HAO
;
Pengyu WU
;
Yanan XIONG
;
Zhiyong LIU
;
Heqin CUI
- Publication Type:Journal Article
- Keywords:
miR-205;
v-yes-1 yamaguchi sarcoma viral oncogene homolog 1;
lung neoplasms;
cell proliferation
- From:
Journal of Jilin University(Medicine Edition)
2014;(3):493-498
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect the expression levels of the miR-205 in lung cancer tissue and A549 cells and its targeted gene YES1 using qRT-PCR and dual fluorescence protein repoter assay system,and to explore the possible mechanism of miR-205 to inhibit the proliferation of lung cancer A549 cells.Methods The expression levels of miR-205 in 10 cases of lung cancer tissue and adjacent normal lung tissue were detected with qRT-PCR.The cell growth curve and colony formation assay were used to determine the proliferation rate of A549 cells after transfected by miR-205 mimics and control mimics.The sequences of YES1 3′UTR (untranslated region)and mutation target sites of YES1 3′UTR were inserted into the plasmid which expressed green fluorescence protein (pcDNA3/EGFP) respectively to construct the green fluorescence protein plasmids of YES1-3′UTR and mut-YES1-3′UTR. There were six groups in the study:YES1-3′UTR, YES1-3′UTR and miR-205 mimics, YES1-3′UTR and control mimics,mut-YES1-3′UTR, mut-YES1-3′UTR and miR-205 mimics, mut-YES1-3′UTR and control mimics;after the plasmids expressed red fluorescent protein (pDsRed2-N1 )were cotransfected into A549 cells,the extracted protein was detected with fluorescence spectrophotometer.Results Compared with adjacent normal lung tissue,the expression levels of miR-205 in lung cancer tissue and A549 cells were decreased (P<0.05 );the proliferation rate of A549 cells in miR-205 mimics group was lower than that in control mimics group (P<0.05). The fluorescence protein expression level in YES1-3′UTR and miR-205 mimics co-transfected group was lower than that in YES1-3′UTR and control mimics co-transfected group, the difference was statistically significant (P<0.01).The number of cell colony formation of A549 cells in highly expressed YES1 group was higher than that in cell control group (P<0.05).Conclusion MiR-205 may inhibit the proliferation of A549 cells through regulating of the expression of YES1 directly.miR-205 and YES1 are potential therapeutic targets for the biological treatment of tumor.
